JPH06505700A - Method for inactivating viruses and sperm with outer membranes - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Background of the invention: Method for inactivating viruses and spermatozoa with outer membranes 1. Field of invention The present invention relates to novel methods and novel products for inhibiting the activity of viruses with outer membranes. do. The present invention also relates to the treatment of viral infections. Invention (to inhibit the onset of diseases and infections caused by viruses with outer membranes) Regarding the method. In one embodiment, the present invention G! In particular, the main mode of infection is sexual The present invention relates to a method for inhibiting the activity of a virus. Write other aspects as well. The present invention provides a method for inhibiting the activity of a virus whose infection mode is a non-sexual outer membrane. Also related. The composition used in the present invention (1) is a virus or virus-infected composition. It is also effective in inhibiting bacteria and fungi that coexist with the disease. Moreover, the present invention Virus-related diseases, details: AIDS, related sexually transmitted infections and other Concerning the treatment of opportunistic infections in immunocompromised hosts.

The method of the invention provides a method for inhibiting viruses and treating viral infections. Concerning the use of mixtures of amines and amine oxides. Betaine is the alkyl of (a) --betaine, alkyl-N-sulfobetaine, acyl-N-betaine, alkyl-N-sulfobetaine, N-substituted aminopropionic acid and alkylimidazolinium betaine. and the amine oxide is selected from the group consisting of (b) alkyl-N.

N-dimethylamine oxide, alkyl-N,N-dihydroxydiethylamine oxide oxide and acylamide 1-lysine and their lower alkyl congeners do.

The term refers to the sulfonic acid analog of such compounds, N-dimethylamino Also includes nopropionic acid and sulfobetatain. Unless otherwise specified, N-jime It represents a chill compound. More specifically, the present invention aims to inhibit viruses with outer membranes. Alkyl N-di(lower alkyl)betaine and alkyl N-di How to use mixtures of (lower alkyl) amine oxides and these viruses Concerning the use of this mixture in the treatment of infections caused by. This mixture is spermicidal It can be used as an agent alone or together with conventional spermicides. These mixtures Compounds can be used in conjunction with contraceptive devices, and can also be incorporated into contraceptive devices. Wear.

2. Description of prior art Certain amine mixtures are effective antimicrobial agents against Gram-negative bacteria and Gram-negative bacteria. It is known to be a drug.

For example, alkyl-N-betaine, alkyl-N-sulfobetaine, acyl-N- betaine, alkyl N-substituted aminopropionic acid or alkylimidazolinium Mubetaine and (b) alkyl-N,N-dimethylamine oxide, alkyl -N, N (dihydroxydiethylamine oxide or acylamide t-amide A mixture of oxides can be used to remove pathogens from the skin, cleanse and deodorize it It is known that (for example, E, B.

U.S. Pat. No. 4,183°952 issued to Michaels; E, B , U.S. Pat. No. 4,075,350 issued to Michaela and E, B.

U.S. Pat. No. 4.107°328 issued to Michael S. (see ). These compositions are also useful for long-term control of body odor.

The same mixture of active ingredients can reduce the oral microflora and inhibit the formation of dental plaque. It is also known to promote oral hygiene. (To E, B, Michaels No. 4,839,158). This type of composition also has It is also known to be an effective fungicide.

Cornet, A. M., et al., AvtimicrobislAgg++ts an d Chea+thetiPL 32(3):350-353 (1988 March), Co rne r, A, M,, The Jo++rnsl o fClinical Denjistt7. Mo1.2(2)+34-38 (1990) Volume 2 (2): 34-38. (1990)). (E, B.

U.S. Patent No. 4,075.350 issued to Michael S. No. 4.107.328, No. 4.183.952 and No. 4 , 839).

Non-ionic with ether or amide bonds between the hydrophilic and hydrophobic parts of the molecule A surfactant that rapidly inactivates the infectivity of herpes simplex virus I and II. It is also known to be sexualized. (Asculai, S.S.

et al., Antimicrobial Agents and Chemotherapy, Vol. 13: 686-690 (1978)) . Asculai et al. also used cell killing activity as an index of antiviral activity. This is clearly stated. Although the composition utilized in the present invention had low cell killing activity, However, this composition inactivated viruses and bacteria. It is advantageous over the composition described by i et al.

The spermicide Nonoxyno l-9 (N-9) has bactericidal and virucidal properties. It is known to be effective.

For example, N-9 is a virus associated with AIDS and herpes simplex virus (H 3V I and II). In-vitro , N-9 can cause gonorrhea, chlamydia, syphilis, trichomoniasis and other sexually transmitted diseases. It has been shown to be effective against the bacteria that causes it. (United States Pharmacopeial Conference USP (See DIUpdate, 1990, pp. 755-756).

Summary of the invention The mixture of betaines defined below inhibits the activity of enveloped viruses. I surprisingly found it to be effective: (a) Alkyl-N-betaine, alkyl-N-sulfobetaine, acyl-N- Betaine, alkyl-N-, a.

betaines, such as minopropionic acid and alkylimidazolinium betaines; and (b) alkyl-N,N-dimethylamine oxide, alkyl-N,N-di such as hydroxydiethylamine oxide or acylamide t-amide oxide. A mixture of amine oxide, carrier and excipient in an acceptable diluent. The present invention Provides a method to inhibit the activity of viruses with outer membranes whose primary mode of infection is sexual do. The present invention provides methods for treating diseases and infections caused by viruses with outer membranes. Regarding. These compositions contain viruses or bacteria that coexist with viral infections. and inhibition of fungi and viruses or fungi and bacteria that coexist with viral infections. It is also effective in treating infectious diseases. Similarly, the present invention relates to a virus having an outer membrane. Provides a method for disinfecting air and surfaces contaminated by.

The present invention also provides a method for preventing enveloped virus infection.

The composition used in the method of the invention can also be used as a spermicide. this Their compositions can be used alone, with other known spermicides, or in combination with contraceptive devices, e.g. Domes, sponges, vaginal inserts, contraceptive films, contraceptive pessaries, suppositories, contraceptives It can also be used by incorporating it into a viscous agent or sustained release device. These compositions are , it can also be integrated into an irrigation system. This composition can be incorporated into a wipe. You can also When used as virucides, these compositions alone with other spermicides and with or incorporated into the contraceptive devices mentioned above. It can be applied. This composition includes foams (IosmL gel, cream, ointment, It can be applied in the form of a jelly or lotion. This composition consists of a liquid, It can also be applied in the form of an aerosol, mist or spray.

Brief description of the drawing Figure 1 shows the effect of C31G (composition according to the invention) on sperm motility and It is a graph showing the effect of Nonoxyno l-9 (N-9).

Figure 2 shows the effect of C31G on total sperm count and Nonoxoyno1- 9 (N-9) is a graph showing the effect.

Figure 3 shows the effect of C31G and Nonoxoy on the average progression rate of spermatozoa. It is a graph showing the effect of no.l (N-9).

Figure 4 shows the toxicity of 031G versus N-9 to 5up-TI cells in vitro. This is the graph shown.

Figure 5 shows a 5-day toxicity test on OEM cells with C31G vs. N-9. This is a graph.

FIG. 6 is a graph comparing the effect on H3V between C31G and N-9.

FIG. 7 is a graph showing the effect of C31G on HIV.

Figure 8 is a graph comparing the inactivation of HIV by C31G and N-9. Ru.

Figure 9 is a diagram comparing the inactivation of HIV by C31G and N-9. .

Detailed description of the invention The method of the present invention involves the inhibition of enveloped viruses and the inhibition of enveloped viruses. Use of mixtures of betaine and amine oxides in the treatment of viral infections Regarding. Similarly, the method of the present invention is a reliable method for inactivating viruses with an outer membrane. Concerning the use of mixtures of amines and amine oxides. Since it is used in the present invention The available betaines include alkyl-N-betaine, alkyl-N-sulfobetaine, and alkyl-N-betaine. syl-N-betaine, alkyl N-substituted aminopropionic acid or alkylimi dazolinium betaine or mixtures thereof. Amines that can be used in the present invention Oxide is alkyl-N,N-dimethylamine oxide, alkyl-N,N- Dihydroxydiethylamine oxide or acylamide t-amide oxide or is a mixture of them. These compositions are effective against viral, fungal and microbial infections. , and is also effective in treating contamination. These compositions can be used to sexually or non-sexually It is effective in inhibiting viruses that have a protective outer membrane. These compositions It is equally effective in inhibiting bacteria and fungi that coexist with or viral infections. this Their compositions can also be used topically.

Typically, these compositions are applied to areas where the virus can infect. This includes Including applications to the vagina, mucous membranes, and skin.

These compositions can be used alone or in conjunction with other spermicides as spermicides. You can also do that. These compositions are suitable for use in contraceptive devices such as condoms, contraceptive pessaries, etc. - Incorporated into sponges, contraceptive films, suppositories, sustained release devices and contraceptive adhesives. You can also use For example, the composition may be used as a lubricant applied to a condom. or incorporated as part of the condom tip reservoir or contraceptive sponge , into contraceptive films, suppositories, sustained release devices or contraceptive adhesives.

The contraceptive film contains the composition described above, type A gelatin, and cellulose rubber. and polyhydric alcohols. The composition is a contraceptive foam, gel, or gel. It can also be used in creams. This composition can also be used in irrigators. can also be used in the prevention of viral infections; these compositions alone can be used with other spermicides or incorporated into contraceptive devices as described above. Ru. The invention also applies to anatomical organs that are exposed to bodily fluids emitted by a partner during sexual activity. Viruses that cause sexually transmitted diseases, consisting of applying the composition to the site. including methods for inhibiting infection.

These compositions are more effective against mammalian cells than Nonoxoyno l-9 (N-9). It was unexpectedly found that the toxicity of the drug was low (see Figures 4 and 5). Table 1 is , it has been shown that these compositions have a spermicidal effect equivalent to that of N-9, and this composition The product is also one of the few surfactants approved by the FDA for use as a spermicide.

When used for topical application, the composition may be used as a liquid, cream, ointment, lotion, It can also be incorporated into foams and gels.

The same (the present invention applies to air in an operating room or examination room, for example, provides a method for disinfecting areas contaminated with enveloped viruses. these The composition can be sprayed, misted, wiped, aerosolized, or sprayed, misted, wiped, aerosolized, or It can also be incorporated into a device that generates files. Using these compositions as liquids You can also

According to the method of the invention, these compositions contain virucides, fungicides and bactericides. It can also be used as a reagent.

The composition for use in the present invention comprises a mixture of betaine and amine oxide. Book Betaines used in the invention include (a) alkyl-N-betaines, alkyl-N-sul fobetaine, acyl-N-betaine, alkyl N-substituted aminopropionic acid, The group consisting of rukylimidazolinium betaines and mixtures of two or more thereof selected from. Typically, betaines contain two lower alkyl atoms attached to a nitrogen atom. It has a group. Most effectively, they have two methyl substituents on the nitrogen atom Ru. The amine oxide used in the present invention is (b) alkyl-N,N-dimethylamine. oxide, alkyl-N,N-dihydroxydiethylamine oxide or alkyl-N,N-dihydroxydiethylamine oxide selected from the group consisting of amine t-amine oxide and mixtures of two or more thereof. selected. Typically, the betaine and amine oxide components are in a molar ratio of 1:5 or 5=1, preferably the molar ratio of alkyl-N-betaine used, alkyl-N- Sulfobetaine, acyl-N-betaine, alkyl N-substituted 2-aminopropio acid and alkylimidazolinium betaine (also known as cocoamphoacetate) ) each have the following structure: or (In the formula, R is a higher grade compound having 10 to 18 carbon atoms, preferably 12 to 166 carbon atoms. It is an alkyl group.

As used herein, the term lower alkyl refers to alkyl having 1 to 3 carbon atoms. Refers to the rukyl group.

Examples of these aforementioned substances include (1) co)-N-betaine, cetyl-N-betaine; Tine, stearyl-N-betaine, isostearyl-N-betaine, oleyl -N-betaine, (2) coco-N-sulfobetaine, cetyl-N-sulfobetaine stearyl-N-sulfobetaine, isostearyl-N-sulfobetaine, Oleyl-N-sulfobetaine, (3) Cocoamide-N-betaine, cetylamide do-N-betaine, stearylamide-N-betaine, isostearylamide- N-betaine, oleyl-amino-N-betaine, (4) N-coco-2 aminop Ropionic acid, N-cetyl-2-aminopropionic acid, N-stearyl-2-amino acid Nopropionic acid, N-isostearyl-2-aminopropionic acid, N-oleyl -2-aminopropionic acid, N-stearyl-bis(2-aminopropionic acid) , N-oleyl-bis(2-aminopropionic acid), N-coco-bis(2-aminopropionic acid) nopropionic acid), N-cetyl-bis(2-aminopropionic acid), (5) N- lauryl-bis(2-aminopropionic acid) 1-hydroxyethyl-1-carbo oxymethyl-2-decylimidazolium betaine, 1-hydroxyethyl-1- Carboxymethyl-2-dodecylimidazolium betaine, 1-hydroxyethyl Ru-1-carboxymethyl-2-cocoimidazolium betaine, 1-hydroxy Ethyl-1-carboxymethyl-2-stearylimidazolium betaine, 1- Hydroxyethyl-1-carboxymethyl-2-olelimidazolium betai or mixtures thereof.

The term “coco” used in this text refers to CTFA (Cosmetic and To Iletryand Fragrance As5ocation.

Washington, D., C. designation), and refers to the alkyls present in coconut oil. represents a mixture of groups, ie alkyl groups of 10 to 18 carbon atoms. in the text The designations of the listed compounds are the same as those of CTFA.

(1) Alkyl-N,N-diethyla used as component (b) of the above mixture. amine oxide, (2) alkyl/l/-N,N-dihydroxylethylamine oxide (3) Acylamide t-amine oxide each has the following structure. do.

(wherein R is a higher alkyl group having 10 to 18 carbon atoms, e.g. Badecyl, undecyl, lauryl, tridecyl, myristyl, cetyl, stearyl oleyl, isostearyl or oleyl. typical amine oxide is decyl-N,N-dimethylamine oxide, lauryl-N,N-dimethylamine oxide, amine oxide, stearyl-N-N-dimethylamine oxide, oleyl-N, N-dimethylamine oxide, Coco N, N-dihydroxydiethylamine oxide , cetyl-N,N-dihydroxydiethylamine oxide, oleyl-N,N-di Hydroxydiethylamine oxide, N,N-dihydroxydiethylamine oxide , oleyl-N-N-dihydroxyethyl-amine oxide and mixtures thereof It is a thing. ) Components (a) and (b) are mixed as usual, then 0. Adjust the pH of the 5% solution from 4 to 8 , preferably the amount of acid necessary to adjust the pH to 4.5 to 5.5. Said The pH of an aqueous solution containing the ingredients of the invention listed in 1. Typically, an aqueous solution of 0.5% of the total weight of active ingredients in the glass electrode is used. Measure using.

Typically, the acid used to adjust the overall composition to the required pH will be All organic or inorganic acids suitable for specific uses, such as hydrochloric acid, phosphoric acid, sulfuric acid, citric acid. acids, such as acetic acid or nicotinic acid. The residue of the composition after using the acid is Usually an acceptable solvent, such as 100 parts or more of water or a lower (C1-04)-valent fat. It is a family alcohol. If water is used, a lower alkyl alcohol, e.g. A small amount of ethanol or propatool may also be added to facilitate formulation. permission An acceptable diluent, solid.

excipients such as ethyl, isopropyl alcohol, polyethylene glycol, etc. Polyhydric alcohol, glycerin, cellulose gums, gelatin, colorants and fragrances. If necessary, the pH of the total acid The required pH is then adjusted by adding a suitable inorganic or organic acid of be able to. The preparations thus obtained contain viruses, bacteria and Substantially homogeneous, uniform and relatively non-toxic compositions with improved activity against fungi It is.

These compositions (1) have low toxicity to mammalian cells and are It is known to be correlated with low irritation and low toxicity when used together (Jou rnal C11nical Dentistry 2 (2): pages 34-38 (1990)), and (2) despite having low cytocidal activity. It was unexpectedly discovered that viruses with outer membranes were inactivated extremely effectively. It was brought out. In the past, cell killing effect was used in research as an index of activity. . (Asculai.

S, S,, Ant 1m1crb, Agents Chemother, 13:6 pp. 78-690 (1978)).

In practice, the total amount of components (a) and (b) in the entire composition will depend on the intended method of use. It can be from 0.01% to 40%, preferably from 0.03% to 30%, depending on the situation. Wear.

For example, the concentrations used are approximately 20% to 30% for contraceptive films and 0% for gels. ．． 2% to 2%.

The composition used in the present invention comprises alkyl N-di(lower alkyl)glycine and alkyl N-di(lower alkyl)glycine and alkyl N-di(lower alkyl)amine oxide, where the lower alkyl is It is 01-3. In the present invention, betaine has been recognized as particularly useful by the present inventor. one of the alkyl N-dimethylbetaines, such as cocobetaine and lauryl Examples include betaine. Amine oxides particularly useful for use in the present invention include alkaline Kyl N-dimethylamine oxide, such as cocodimethylamine oxide and la It is uryldimethylamine oxide.

One particular composition that can be used in the present invention includes cocobetaine, cocoa oxide and citric acid monohydrate.

Another composition that can be used in the present invention is lauryl betaine, lauramine citric acid and citric acid-hydrate.

In such compositions, the molar ratio of betaine to amine oxide is typically 5:1. to 1:5, preferably a molar ratio of 2:1 to 1:2, more preferably about 1=1. Ru.

Other compositions that can be used in the present invention include betaine, amine oxide, Blo om strength 100 to 300 and molecular weight about 75.000 to about 300.000 of gelatin, polyhydric alcohol and cellulose gum. like this The compositions are filed concurrently with the present application, U.S. Patent Application Serial No. 07/673,6. No. 31, co-pending US patent application Ser.

The composition used in the present invention inhibits the activity of viruses associated with AIDS. Can be done. These compositions are capable of inhibiting H3V-1 and H3V-2 activity. It is also expected that The composition used in the present invention is ASB.

It is also expected that it can inhibit hepatitis C. This composition is suitable for sexually transmitted diseases (S). TD) can inhibit viruses, bacteria and fungi associated with He is also expected to be deaf.

Compositions of the invention may be useful in connection with: 1) HIV infection is often accompanied by other viral and/or microbial pathogens. Often associated with infection. In fact, some researchers believe that HIV causes AIDS. suggested that it may not be the only pathogen (Duesberg , P.H. (1991) proc.

Natl. Acad. Sci. 88:1575-1579;, 2Lemaitr. e, M., Guetard, D.;

Hen1n, Y., Montagnier, L., and Zerial, A. ( 1990), Res, Virol.

141: see 5-16). For this reason, for example, the Has a broad spectrum of activity against viruses, bacteria, and yeasts such as Kanjig. Antimicrobial agents are used in the prevention and treatment of acquired immunodeficiency syndrome (AIDS). It may be particularly important.

2) Certain bacteria known to cause STDs may facilitate HIV infection. It is thought to be long.

In people exposed to HIV, the bacteria that cause STDs are often other They do not respond to treatments that are very effective for people with cancer. HIV infection is It may promote the transmission of bacterial STDs, which in turn may promote the transmission of HIV.

STDs, such as chlamydia, genital sores, syphilis, genital herpes, and gonorrhea, can cause skin ulceration and the risk of sexually acquiring or transmitting HIV infection. It is thought to increase sex. Aral, S.

00. 5 scientific American.

264 (2): pp. 62-69 (February 1991).

The compositions may also be used to treat other enveloped viruses, such as vaccinia, varicella, zoster. herpes, cytomegalovirus, EB virus, influenza, mumps Also beneficial in inactivating hypodenitis, measles, rhinovirus, rabies and rubella There are cases. For a deeper understanding of the invention, the following examples are mainly used in further detail. A specific example is provided for illustrative purposes. The present invention is as described below in the claims. The composition contains C31G, cocobetaine and cocodimethylamine oxy (CTFACosmetic and Toiletry andFragr ance As5ocjationS Washington, D.C.) [CFTA] It is a concentrate of equimolar preparations of and can be used in many different forms.

Cocobetaine; 31.5% Active Ingredient (AI): 183, 9 k g (40 5,8 bond), cocoa amine oxide, 31.5% (AI): 147, 4k g (325 lbs.), citric acid-hydrate, USP: 11.8 kg (26 lbs.) ), Purified Water, USP: 26 lbs. at 29.6% AI (7) C31G to 354. 9 k g (782,5 lb) made, 1% AI diluted; pH=4.9° 1 The spermicidal action of C31G in Example 1 was evaluated using nonoxyn o1-9 (N-9 ) was compared with the spermicidal effect of

This test was conducted using a Hami 1 ton-Thorn sperm motility analyzer. . A collected and washed semen sample was diluted with the two compounds and incubated for 15 seconds. (After testing, make three identical copies for analysis), and then perform the best inactivation process according to the criteria below. Small concentrations were measured; 1. Inhibition of sperm motility (Figure 1) 2. Decrease in total sperm count (Figure 2) 3. Inhibition of average sperm progression rate (Figure 3) As shown in Table 1, C31G and N- 9 obtained the same result.

Table 1 Minimum concentration for spermicidal action C31G N-9 Total inhibition of motility 150ppm 90% in 150ppm sperm count Reduction of 150 pI) m 50% at 150 ppm progression rate decrease 125ppm 125ppm to Several types of assays for the cytotoxic effects of 031G and N-9 on mammalian cells It was measured by Cytotoxicity occurs when surfactants are used in contraceptives or prophylactic drugs. as an indicator of relative safety and comfort.

As shown in Figure 4, the drug effect on 5UP-TI cells (human lymphocyte cell line) was observed. In a two-week test, the same concentration of N-9 was extremely toxic, while C 31G showed minimal toxicity at 0.001% [10 ppm].

bile Figure 5 shows MMT reduction in OEM cells after 5 days of continuous exposure. It is. Under these conditions C31G ft, 3ppm c, 0003%] N-9 is not toxic to mammalian cells at all, whereas N-9 is toxic at the same concentration. , reduced mammalian cell viability by 50%.

A compound that has the same spermicidal effect as N-9 is less toxic to mammalian cells than N-9. The low level is unexpected and indicates that C31G's antiviral effect is lower than that of HSV-1. and HIV-1.

Testing the effects of 031G and N-9 on herpes simplex virus type 1 (K2S strain) To test the virus, we injected the stored virus into Vero (African green monkey kidney) cells. Prepared. The virus was released from the cells by one freeze-thaw cycle and then Sonication was performed. When this virus titer was measured using Vero cells, it was 4.5X10 The 8 plaques were the same as the drug in the spermicidal test.

The stock concentration of each drug was 5%. Make 2x serial dilutions and adjust pH in PBS at room temperature. 7,41:OirO, diluted to 04%. Dilute 24 μl of virus 22 μl each time. 0 μl (108 PFU) was added to give a final concentration of 0.5 to 0.004%. child Incubate these samples at room temperature for 10 minutes, then add 100 ul of each immediately. Immediately, it was diluted to 1 ml with ice-cold DMEM 15% FBS. Then serially dilute 10 times of remaining infectious virus on Ver0 cells in 24-well plates. The titration was lowered to 10. Plaques were counted 36 hours after infection.

Table 2 Concentration (%) Remaining virus (PFU/ml) N-9C31G O,0043,5X108 3.5X1080.008 2.7X108 3. 3X1080.015 1. lX108 2.5X1050.03 3200 <167 0.06 <167 <167 0.13 <167 <167 0.25 <167 <167 0, 5 <167 <167 These are titers (i.e. PEU/mL) rather than absolute numbers of PFU. I would like to draw your attention to this. The titer measured on untreated virus was the same as in the experiment using N-9. 4.7 x 108 PFU/mL, and 4.5 x 108 P in experiments using C31G. It was FU/ml.

Complete inhibition of plaque formation by C31G requires more than one dilution less than N-9 Notice what happens at a point in time. Please refer to FIG.

Example 6 Inactivation of the HIV-1 (AIDS) virus was studied in two experiments. first try In the experiment, the effect of 031G on HIV-1 was evaluated at two different exposure times, 2 minutes and 031G. Comparisons were made at 45 minutes and 45 minutes. In the next study, C31G and N-9 against HIv. The same is true for the relative effects.

virus strain, and the virus titer was determined by the reduction of P-24 HIV antigen. The antiviral activity was measured and compared.

The protocol and results of the first study are as follows.

Effect of C31G on human immunodeficiency virus Materials: Virus: HIV-i strain 3B (5XIOID50 units/ml) Cells: 5UP-'rt cell line (giant cells and syncytial cells when infected with HIV) C D 4 + + J lymphocyte-like cells that produce unique cell lysis with Cytium Rotocol: 1. C31G was prepared in PBS with serial 4-fold dilutions (4.0% 0.004%, and PBS alone as a control). pH is 5.5 It was adjusted.

2. Prepare a certain amount of stored virus, add 031G, detergent, and virus ratio. Each concentration was added at a concentration of 1:9 (1:10 detergent dilution). These are 2 minutes or 45 Incubated for minutes.

3. Each virus/cleaning agent diluted to 1:4096 by serial 4-fold dilution A mixture was prepared.

4. 16.6 μl of each virus/detergent dilution from each round was added to 5UP-TI cells. Two replicate wells (104 cells in 150 μm volume: detergent dilution 1:10) diluted 4 times).

5. Test the wells twice a week for two weeks, with each well containing a unique Regarding the presence of syncytia of viruses, or non-syncytia due to bacteriolysis of detergents. The results were evaluated as positive or negative for mucosal ghosting.

note: Virus was added to C31G at a 1:10 dilution of the initial concentration for either 2 or 45 minutes. exposure for a period of time, then another 1:10 dilution, and then a series of 4-fold dilutions for the entire period. Ta.

Cells were incubated in C31G at 1:100 and then in serial 4-fold dilutions of the initial concentration for the entire period. Exposed.

See Figure 7.

Example 7 The protocol and results of the second study are as follows: material: Cell: CEM cell line ATCC #119T cell Virus: HI V I I I b Test compounds: Compounds C31G and N-9 supplied as 5% solutions were diluted Compounds in PBS were serially diluted 1 and 2 times before passing through a low binding 0.22 filter. Samples N-9 and C31G were adjusted to pH 5.5. The highest concentration is 4%, the lowest is It was 0.03%. The initial concentration is 4.2.1.0.5.0.25.0.125 ．． 0.06 and 0.03%, similar to the no-drug control.

Control titrations were performed twice.

2.14 Each concentrated virus in a 12x75mm sterile plastic tube (minimum lXl0TCID/ml) was prepared. Add 0.1 ml of each compound dilution. (1=10 dilution of initial drug concentration).

3. Incubate for 10 min at room temperature and in complete growth medium (on ice). Completed (or delayed) by diluting 1:10.

4. Additional serial 5-fold dilutions (final dilution 1:78) in complete medium on ice. 7 dilutions equal to 1250) were prepared.

5. Add 50 μl of virus dilution to a 96-well tray (U-bottom) 4 times. 3 x 104 CEM cells in 50 μl were added to the wells.

6. Allow virus to be absorbed for 60 minutes. The cells were plated at 150 Orp. Washed twice by centrifuging at m for 5 minutes and aspirating the supernatant. final precipitation Cells were resuspended in 100 ul of medium and transferred to a 96 flat-bottom well tray. this The results are shown in FIGS. 8 and 9.

Example 8 Prescription of contraceptive/infection prevention products Concentrate (CCon) is a mixture containing: Lauryl betaine (30% at) 1000 parts lauryl betaine (30% ai ) 870 parts Citric acid monohydrate 69 parts Stir the above to make a homogeneous solution. When diluted 30 times, this composition The pH is set to 4.85. Estimated concentration equals 28.5% active ingredient (ai) .

Example 9 Suppositories (inserts) used with or without condoms CCon (Example 8) 526 copies PEG-32 4000 copies PEG-201000 copies Povidone 74 parts The above is stirred to form a solution at 45°C, poured into molds, cooled, taken out and packaged.

Example 10 contraceptive sponge CCon (Example 8) 3.5gm5 Put into each mold to make cured urethane. child The mold of the cervix is fitted with a polyester loop for removal from the sponge after intercourse. Along with lids, condoms, and contraceptive pessaries.

or spermicidal gel, cream or jelly used alone before intercourse.

Gel-I CCon (from Example 8) 7.0 parts Gelatin A200 1.5 parts Glycerin 10.0 parts hydroxyethyl cellulose (High viscosity product) 0.4 part Water 81.1 parts Gel - 23, 5 parts CCon (from Example 8) 1, Opt Nonoxyno Same as above except for substituting l-9.

Procedure - Gelatin and cellulose gum are ground in glycerin and added to water at 45°C. , stir to dissolve. Add the surfactant to the container, remove the warm solution and package it. Ru. When cooled, it forms a homogeneous, fluid, highly viscous gel.

Jelly 1 and 2 Clear liquid contraceptive jelly is made from gelatin and hydroxyethyl cellulose as described above. 1.5 parts of high viscosity hydroxypropyl or hydroxypropyl methyl Prepared by substituting ululose.

cream 1 and 2 The above gel formulation consists of dissolving 0.5 parts of cetyl alcohol in glycerin at 49°C. and add it to the gel 1 or 2 formulation before grinding the gelatin and cellulose gum. It can be converted by matching.

Example 12 contraceptive film 6.2 pounds of type A gelatin is mixed with 0.5 pounds of hydroxyethyl cellulose. Crush together. 31 Add pontonoglycerin. Mix the solution thoroughly to create a slurry. to form. Add 33 bonds of CCon from Example 8 and 15 pounds of water. 40 Warm to ℃. Mix until gum and gelatin are fully hydrated. The solution is A film with a thickness of approximately 3 mm is cast on an ethylene sheet, and after cooling, the film is Cut it out and use it as a contraceptive film.

9yo0LX-4becct:in proof 9yo OLX Yumi-Sumi 9yo OLX 薯渑-稠 ±1■ 4N0L911 Kouko Sound 6-9r -6J-6't10-Cφ back cup international search report +m Pass WI Am1tell # NaρCT/US 91105060! ,@, 1. el　Aemlct11a++++SayaPσYaba91105060------^- ---・　ρCT/us　91105060 International Search Report PCTM 91105060 S^　4%17

Claims (19)

[Claims] 1. Mixture of betaine and amine oxide at the location of virus with outer membrane inhibiting the activity of an enveloped virus, consisting of applying a composition containing A method, wherein the betaine is an alkyl-N-betaine, an alkyl-N-supra Rufobetaine, acyl-N-petaine, alkyl N-substituted aminopropionic acid or or alkylimidazolinium betaine or a mixture of two or more thereof. selected and said amine oxide is alkyl-N,N-diethylamine oxide , alkyl-N,N-dihydroxyethylamine oxide or acylamide t - selected from amine oxides or mixtures of two or more thereof; How to do it. 2. A composition comprising a mixture of betaine and amine oxide during sexual activity. Sexually transmitted disease prevention, consisting of application to parts of the body that are exposed to body fluids emitted by the other person. A method for inhibiting infection of a causative virus, wherein the betaine is an alkyl -N-betaine, alkyl-N-sulfobetaine, acyl-N-petaine, alkyl-N-sulfobetaine, Kyl N-substituted aminopropionic acid or alkylimidazolinium betaine or is selected from mixtures of two or more thereof, and said amine oxide is an alkyl- N,N-diethylamine oxide, alkyl-N,N-dihydroxyethylamine amine oxide or acylamide t-amine oxide or a mixture of two or more thereof. A method, characterized in that the compound is selected from a compound. 3. A composition containing a mixture of betaine and amine oxide is applied to the site of viral infection. A method for the treatment of viral infections comprising applying betaine to Ki, Ru-N-betaine, Alkyl-N-sulfobetaine, Acyl-N-betaine , alkyl N-substituted aminopropionic acid or alkylimidazolinium betai or a mixture of two or more thereof, wherein said amine oxide is selected from Kyl-N,N-diethylamine oxide, alkyl-N,N-dihydroxyethyl amine oxide or acylamide t-amine oxide or two or more thereof A method, characterized in that the method is selected from a mixture of 4. Using a composition containing a mixture of petaine and amine oxide as a spermicide A method of inhibiting sperm activity by -Betaine, alkyl-N-sulfobetaine, acyl-N-betaine, alkyl N-substituted aminopropionic acid or alkylimidazolinium betaine or the like selected from a mixture of two or more thereof, and the amine oxide is selected from a mixture of two or more thereof, and the amine oxide is N-diethylamine oxide, alkyl-N. N-dihydroxyethylamine oxide or acylamide t-amine oxide or mixtures of two or more thereof A method, characterized in that the method is selected from: 5. A contraceptive device having a composition containing a mixture of betaine and amine oxide. So, this betaine is alkyl-N-betaine, alkyl-N-sulfobetaine , acyl-N-betaine, alkyl N-substituted aminopropionic acid or alkyl selected from imidazolium betaine or a mixture of two or more thereof; amine oxide is alkyl-N,N-diethylamine oxide, alkyl-N , N-dihydroxyethylamine oxide or acylamide t-amineoxy or a mixture of two or more thereof. 6. Use of a composition comprising a mixture of betaine and an amine oxide, Betaine is alkyl-N-betaine, alkyl-N-sulfobetaine, acyl- N-betaine, alkyl N-substituted aminopropionic acid or alkylimidazoli amine betaine or a mixture of two or more thereof; oxide is alkyl-N,N-diethylamine oxide, alkyl-N,N-dihydryl Droxyethylamine oxide or acylamide t-amine oxide or the like A virus having an outer membrane, characterized in that it is selected from a mixture of two or more of these. Use for the preparation of drugs that inhibit activity or are used as spermicides. 7. Treating viral infections comprising a mixture of betaine and amine oxide , wherein the betaine is an alkyl-N-betaine, an alkyl-N-supra Rufobetaine, acyl-N-petaine, alkyl N-substituted aminopropionic acid or or alkylimidazolinium betaine or a mixture of two or more thereof. and the amine oxide is an alkyl-N,N-diethylamine oxide, Alkyl-N,N-dihydroxyethylamine oxide or acylamide t- selected from amine oxides or mixtures of two or more thereof , medicine. 8. The composition comprises alkyl N-di(lower alkyl)betaine and alkyl N-di( lower alkyl) oxide, wherein the lower alkyl is C1-C3. The method or apparatus according to any one of items 1 to 7. 9. Alkyl N-di(lower alkyl) betaine and alkyl N-di(lower alkyl) claim 1, wherein the molar ratio of the amine oxide (kill) amine oxide is from 1:5 to 5:1. 9. The method or apparatus according to any one of Item 8. 10. The composition contains alkyl N-di(lower alkyl) betaine, alkyl N-di(lower alkyl) betaine, alkyl N-di(lower alkyl) alkyl) amine oxide, and water as a carrier that is acceptable to the pH of the composition. The claimed invention contains an amount of acid sufficient to adjust from 4 to 8 when measured in aqueous solution. 9. The method or apparatus according to any one of paragraphs 1 to 9. 11. The method or apparatus according to any one of claims 1 to 10. where the composition includes: a) alkyldimethylglycine, b) an alkyldimethylamine oxide and c) the ingredient in an acceptable carrier. The pH of the composition when measured in aqueous solutions of (a) and (b) is between 4 and 8. sufficient amount of acid to adjust to. 12. Alkyl N-di(lower alkyl)betaine is cocobetaine or uryl - petaine and the alkyl N-di(lower alkyl)amine oxide is cocoa amine. of claims 1 to 10, which is lauramine oxide or lauramine oxide. A method or apparatus according to any one of the preceding paragraphs. 13. The composition may be applied to mucous membranes, skin, vagina, inert surfaces, or dispersed in the air. 7. The method according to any one of claims 1, 2, 3, 4 or 6. 14. Betaine and amine oxide compositions can be used as foams, gels, creams, ointments, In the form of a jelly, lotion, liquid, spray, mist, aerosol or wipe Any one of claims 1 to 13, blended or applied in The method or apparatus described in Section. 15. The virus is HIV, AIDS-related virus, HSVI, HSVII, A Hepatitis, hepatitis B, hepatitis C, vaccinia, chickenpox, herpes zoster, cytomegalo viruses, Epstein-Barr virus, influenza, mumps, hemp Viruses associated with measles, rhinovirus, rabies or rubella; The method according to any one of items 1 to 3, or item 6 or 7. 16. Claims that the composition is used with or incorporated into a contraceptive device The method according to any one of paragraphs 1 to 4 and 6 to 12. 17. The composition is incorporated into a lubricant applied to a condom, or the composition is applied to a condom. The composition is applied to contraceptive sponges, contraceptive films, and Claim 5, incorporated into a release device, suppository, foam, gel, cream or jelly. Contraceptive devices as described in . 18. An irrigator comprising a mixture of betaine and an amine oxide, the irrigator comprising: The betaines listed above are alkyl-N-betaines, alkyl-N-sulfobetaines, acyl -N-betaine, alkyl N-substituted aminopropionic acid or alkylimidae an amine selected from zolinium betaine or a mixture of two or more thereof; oxide is alkyl-N,N-diethylamine oxide, alkyl-N,N- dihydroxyethylamine oxide or acylamide t-amine oxide or is selected from a mixture of two or more thereof. 19. A wiper comprising a mixture of betaine and amine oxide, The betaines listed above are alkyl-N-betaines, alkyl-N-sulfobetaines, acyl -N-betaine, alkyl N-substituted aminopropionic acid or alkylimidae an amine selected from zolinium betaine and mixtures of two or more thereof; oxide is alkyl-N,N-diethylamine oxide, alkyl-N,N- dihydroxyethylamine oxide or acylamide t-amine oxide or is selected from a mixture of two or more thereof.